scholarly journals Investigating the mechanisms of indocyanine green (ICG) cellular uptake in sarcoma

2021 ◽  
Author(s):  
Corey D Chan ◽  
Marcus J Brookes ◽  
Riya Tanwani ◽  
Chloe Hope ◽  
Toni A Pringle ◽  
...  
Keyword(s):  
Indocyanine Green ◽  
Cell Line ◽  
Cell Lines ◽  
Cellular Uptake ◽  
Incubation Time ◽  
Near Infrared ◽  
Cancer Cell Line ◽  
Proliferation Rate ◽  
Low Grade

Introduction: Indocyanine green (ICG) is a near infrared (NIR) dye which has been used clinically for over 50 years and has recently been utilised for fluorescence guided surgery in a number of cancer types, including sarcoma. ICG is taken up and retained by sarcoma tumours to a greater extent than normal tissue, demonstrating its potential to aid in visualisation of tumour margins. However, the mechanisms surrounding preferential ICG uptake in tumours are poorly understood. Methods: In vitro ICG cellular uptake studies were performed across a panel of four sarcoma cell lines and one breast cancer cell line, exhibiting varying proliferation rates and phenotypes. The effects of ICG concentration, incubation time, inhibition of clathrin mediated endocytosis and cell line proliferation rate on the cellular uptake of ICG were investigated, using fluorescence microscopy and flow cytometry. The spatial orientation of ICG was also assessed in a patient specimen. Results: The level of ICG cellular uptake was dependent on ICG concentration and incubation time. Cell line proliferation rate correlated significantly to ICG uptake within 30 minutes (Rs = 1, p<0.001), whilst retention of ICG after 24hrs did not (Rs = 0.3, p=0.624). From our data, the primary mechanism of ICG uptake in sarcoma cells is via clathrin mediated endocytosis. Following the resection of a grade 3 leiomyosarcoma, ICG signal was detectable macroscopically and on 3um sections, whilst being negative on the muscle control. Conclusions: The use of ICG for tumour detection in sarcoma surgery may demonstrate higher utility in high grade tumours compared to low grade tumours, due to the observation of higher ICG uptake in more proliferative cell lines. It is likely that the enhanced permeability and retention (EPR) effect plays a significant role in the retention of ICG within tumours. Future work on the detection of ICG at the cellular level within human tissue sections is required, with the aid of purpose built NIR microscopes.

Design, Synthesis, Molecular Docking, and Anticancer Evaluation of Pyrazole Linked Pyrazoline Derivatives with Carbothioamide Tail as EGFR Kinase Inhibitors

2020 ◽  
Vol 21 (1) ◽  
pp. 42-60
Author(s):  
Farah Nawaz ◽  
Ozair Alam ◽  
Ahmad Perwez ◽  
Moshahid A. Rizvi ◽  
Mohd. Javed Naim ◽  
...  
Keyword(s):  
Molecular Docking ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Line ◽  
Cancer Cell Lines ◽  
Kinase Assay ◽  
Cervix Uteri ◽  
Egfr Kinase

Background: The Epidermal Growth Factor Receptor (known as EGFR) induces cell differentiation and proliferation upon activation through the binding of its ligands. Since EGFR is thought to be involved in the development of cancer, the identification of new target inhibitors is the most viable approach, which recently gained momentum as a potential anticancer therapy. Objective: To assess various pyrazole linked pyrazoline derivatives with carbothioamide for EGFR kinase inhibitory as well as anti-proliferative activity against human cancer cell lines viz. A549 (non-small cell lung tumor), MCF-7 (breast cancer cell line), SiHa (cancerous tissues of the cervix uteri), and HCT-116 (colon cancer cell line). Methods: In vitro EGFR kinase assay, in vitro MTT assay, Lactate dehydrogenase release, nuclear staining (DAPI), and flow cytometry cell analysis. Results: Compounds 6h and 6j inhibited EGFR kinase at concentrations of 1.66μM and 1.9μM, respectively. Furthermore, compounds 6h and 6j showed the most potent anti-proliferative results against the A549 KRAS mutation cell line (IC50 = 9.3 & 10.2μM). Through DAPI staining and phase contrast microscopy, it was established that compounds 6h and 6j also induced apoptotic activity in A549 cells. This activity was further confirmed by FACS using Annexin-V-FITC and Propidium Iodide (PI) labeling. Molecular docking studies performed on 6h and 6j suggested that the compounds can bind to the hinge region of ATP binding site of EGFR tyrosine kinase in a similar pose as that of the standard drug gefitinib. Conclusion: The potential anticancer activity of compounds 6h and 6j was confirmed and need further exploration in cancer cell lines of different tissue origin and signaling pathways, as well as in animal models of cancer development.

scholarly journals Synthesis and Experimental Validation of New Designed Heterocyclic Compounds with Antiproliferative Activity versus Breast Cancer Cell Lines MCF-7 and MDA-MB-231

2017 ◽  
Vol 2017 ◽  
pp. 1-10 ◽  
Author(s):  
Vincenza Barresi ◽  
Carmela Bonaccorso ◽  
Domenico A. Cristaldi ◽  
Maria N. Modica ◽  
Nicolò Musso ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Anticancer Agents ◽  
Cancer Cell Line ◽  
Breast Cancer Cell Lines ◽  
Design And Synthesis ◽  
Human Breast Cell ◽  
Mcf 7

Recent drug discovery efforts are highly focused towards identification, design, and synthesis of small molecules as anticancer agents. With this aim, we recently designed and synthesized novel compounds with high efficacy and specificity for the treatment of breast tumors. Based on the obtained results, we constructed a Volsurf+ (VS+) model using a dataset of 59 compounds able to predict the in vitro antitumor activity against MCF-7 cancer cell line for new derivatives. In the present paper, in order to further verify the robustness of this model, we report the results of the projection of more than 150 known molecules and 9 newly synthesized compounds. We predict their activity versus MCF-7 cell line and experimentally verify the in silico results for some promising chosen molecules in two human breast cell lines, MCF-7 and MDA-MB-231.

PKC412 Shows Strong Anti-myeloma effects in in-Vitro-Studies

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 5172-5172
Author(s):  
Ahmet H Elmaagacli ◽  
Michael Koldehoff ◽  
Nina K Steckel ◽  
Dietrich Beelen
Keyword(s):  
Multiple Myeloma ◽  
Mrna Expression ◽  
Cell Line ◽  
Cell Lines ◽  
Interleukin 6 ◽  
Proliferation Rate ◽  
In Vitro Studies ◽  
Apoptosis Rate ◽  
Rpmi 8226

Abstract Background. The protein kinase C (PKC) inhibitor PKC412 (N-benzylstaurosporine) is a derivate of the naturally occurring alkaloid staurosproine and has been shown to inhibit the conventional isoforms of PKC (alfa, beta1, beta2 and gamma). PKC412 has been shown to have an antitumor effect on non-small cell lung cancer and acute leukemia with FLT3 mutations, but little is known about its effect on multiple myeloma up to date. Methods. Since PKC is also an inhibitor of a tyrosin kinase which is associated with VEGF, and inhibits the release of Interleukin-6, TNF alfa, and that of growth factor dependent C-FOS, we postulated that PKC412 might have also strong anti-myeloma features. Here we evaluated the anti-myeloma effect of PKC412 in the multiple myeloma cell lines INA-6, OPM-2 and RPMI 8226 by measuring its effect on their proliferation rate, the apoptosis rate and the Interleukin-6 mRNA expression. Results. PKC412 showed strong anti-myeloma effects in all three celllines. 50nM of PKC412 was enough to drop the proliferation rate in all three cell lines under 10% compared to untreated cells(p&lt;0.01). The apoptosis rate increased in INA cell line up to 2,5 times and in RPMI cell line up to 3 times (p&lt;0.05), whereas only a moderate increase was observed in the OPM2 cell line with 500nM of PKC412. As expected, the IL-6 mRNA expression decreased after PKC412 treatment in all three cell lines more than 50%. The addition of Bevacizumab to PKC412 in RPMI and OPM-2 cell lines did not increased the apoptosis rate significantly, whereas the addition of short-interference RNA (RNAi) against VEGF increased the apoptosis rate in RPMI 8226 cells about 20% (p&lt;0.05) and in OPM-2 cells up to 30% (p&lt;0.01) compared to PKC412 alone, which was also associated concordantly with a further reduction of the proliferation rate in RPMI and OPM-2 cells up to 30%. Conclusions. PKC412 shows strong anti-myeloma effects and might be effective also in the treatment of patients with multiple myeloma. These in-vitro studies might encourage to initiate clinical trials with PKC412 in patients with multiple myeloma.

scholarly journals PCNA/cyclin expression and BrdU uptake define different subpopulations in different cell lines.

1991 ◽  
Vol 39 (1) ◽  
pp. 23-30 ◽  
Author(s):  
M D Coltrera ◽  
A M Gown
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Proliferation Rate ◽  
Brdu Incorporation ◽  
Cell Populations ◽  
Ki 67 ◽  
Tissue Specimens ◽  
Proliferating Cell ◽  
Two Populations

Monoclonal antibodies (MAb) to a 36 KD protein, proliferating cell nuclear antigen (PCNA/cyclin), have been previously shown to be capable of identifying proliferating cells in vitro as well as in alcohol-fixed, paraffin-embedded tissue specimens. The routine use of these anti-PCNA/cyclin MAb in investigative studies and in diagnostic pathology requires a clearer understanding of the distribution of PCNA/cyclin in the different cell populations found in tissue specimens. We therefore compared the ability of MAb to three nucleus-associated proliferation markers (MAb 19A2 to PCNA/cyclin; Ki-67 to an undefined proliferation-related marker; BU-1 to 5'-bromodeoxyuridine (BrdU) incorporated into DNA) to identify the proliferating cell fraction of various cells in vitro. The cell lines were chosen to represent a spectrum of proliferation rates (high to low) and cell lineage (mesenchymal vs epithelial, non-transformed vs malignant): (a) HeLa and A-431 (two malignant carcinoma cell lines with high proliferation rates); (b) SK-5 (a non-transformed fibroblast cell line with a low proliferation rate); (c) HUVE (a non-transformed human umbilical vein endothelial cell line with a low proliferation rate). Single and double labeling immunofluorescence studies were performed after uniform 1-hr incubations with BrdU. Comparison of the overlapping distributions of detectable PCNA/cyclin expression and BrdU incorporation demonstrated substantial qualitative and quantitative differences between the different cell lines. In two of the four cell lines (HeLa, A-431) the BrdU staining distributions formed inclusive subsets of the PCNA-positive cell populations. In the HUVE cell line the two populations overlapped incompletely. In one cell line, SK-5, the two populations were mutually exclusive. MAb Ki-67 demonstrated a pattern in the SK-5 cell line that was strongly predictive of PCNA positivity, while showing no associated patterns in the other three cell lines. We conclude that PCNA/cyclin expression detected by MAb may define different cell subpopulations in different cell types relative to those incorporating BrdU or expressing the target antigen for Ki-67. This has implications for the clinical study of mixed cell populations using these antibodies.

scholarly journals Cflar Splicing SNP As Predictor of Chemo-Sensitivity to Anticancer Drugs

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 2056-2056
Author(s):  
Lata Chauhan ◽  
Emilie J Bergsma ◽  
Jatinder K Lamba
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Anticancer Drugs ◽  
Cancer Cell ◽  
Cancer Cell Line ◽  
Chemotherapeutic Agents ◽  
Extrinsic Pathway ◽  
Wide Range

Abstract Background: Anticancer therapeutics leverages activation of apoptosis signal transduction pathways (extrinsic and intrinsic apoptotic pathways) in cancer cells. Apoptosis induced by the extrinsic pathway complements that induced by the intrinsic pathway, so targeting extrinsic pathway is considered a useful new therapeutic approach. Preclinical data suggests TNF related apoptosis inducing ligand (TRAIL) as a promising approach as apoptosis of tumor cells is achievable in vivo without lethal toxicities. CASP8 and FADD-like apoptosis regulator (CFLAR) is an inhibitor of death receptor signaling that inhibits TRAIL-mediated caspase 8 auto-activation and subsequent apoptosis. We recently identified a splicing single nucleotide polymorphism (SNP) rs10190751 G>A in CFLAR, where presence of the variant allele (A) was associated with alternate splicing as well as with chemo-sensitivity to chemotherapeutic agent triptolide. However role of CFLAR and the splicing SNP on chemo-sensitivity to wide array of anticancer drugs is not known. Objective: Given the central role of CFLAR in apoptotic pathway, the goal of this study was to investigate impact of CFLAR and its splicing SNP on cytotoxicity of wide range of chemotherapeutic drugs including the ones extensively used in hematological malignancies. Methods: We selected chemotherapeutic agents with wide range of mechanisms of action as blocking DNA biosynthesis, interfering with structure or function of DNA or protein synthesis, interfering with DNA transcription or replication as well as drugs that are cell cycle specific or not. We selected nine Epstein-Barr-virus transformed lymphoblastoid cell lines (LCLs) that are part of International HapMap project representing different genotype for rs10190751 (CFLAR splicing polymorphism; 3 in each genotype category) with twelve different chemotherapeutic agents. Further validation of CFLAR's role in in vitro chemosensitivity was evaluated using CFLAR knockdown and overexpression studies in pancreatic and leukemic cell lines such as Panc-1 and THP1. Results: CFLAR splicing SNP rs10190751, was associated with in vitro cytotoxicity of several chemotherapeutic agents (Bortezomib, SAHA, doxorubicin, sorafenib). The results of screening of 122 FDA approved drugs and their relation with CFLAR as well as its splicing SNP will be presented at the annual meeting. As an example we show below that knock down of CFLAR isoforms have a significant impact on in vitro chemosensitivity to bortezomib and SAHA (Figure 1) Conclusion: Our results suggest critical role of CFLAR in anticancer drug mediated cell death. Additionally splicing SNP in CFLAR seems to play an important role in drug sensitivity/resistance. Therapeutic strategies to directly or indirectly inhibit the expression and/or function of CFLAR might be an attractive option to overcome resistance to wide range of chemotherapeutic agents. Figure 1. Impact of siRNA mediated knockdown or of CFLAR on Bortezomib and SAHA sensitivity in THP1 and Panc-1 cancer cell line. Figure 1. Impact of siRNA mediated knockdown or of CFLAR on Bortezomib and SAHA sensitivity in THP1 and Panc-1 cancer cell line. Disclosures No relevant conflicts of interest to declare.

Finasteride dose-dependently reduces the proliferation rate of the LnCap human prostatic cancer cell line in vitro

Urology ◽  
1995 ◽  
Vol 45 (2) ◽  
pp. 282-290 ◽  
Author(s):  
Mauro Bologna ◽  
Paola Muzi ◽  
Leda Biordi ◽  
Claudio Festuccia ◽  
Carlo Vicentini
Keyword(s):  
Cell Line ◽  
Cancer Cell ◽  
Prostatic Cancer ◽  
Cancer Cell Line ◽  
Proliferation Rate

scholarly journals Antiproliferative withanolides from the Solanaceae: A structure–activity study

2012 ◽  
Vol 84 (6) ◽  
pp. 1353-1367 ◽  
Author(s):  
Huaping Zhang ◽  
Abbas K. Samadi ◽  
Mark S. Cohen ◽  
Barbara N. Timmermann
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Antiproliferative Activity ◽  
Cancer Cell Line ◽  
Human Head ◽  
Human Cell Line ◽  
Withaferin A ◽  
Relationship Analysis ◽  
Structure Activity

As part of our search for bioactive compounds from plant biodiversity, 29 withanolides were recently isolated from three members of the Solanaceae:Physalis longifolia,Vassobia breviflora, andWithania somnifera. Six derivatives were prepared from these naturally occurring withanolides. All compounds were evaluated for in vitro antiproliferative activity against an array of cell lines [melanoma cell lines (B16F10, SKMEL28); human head and neck squamous cell carcinomas (HNSCC) cell lines (JMAR, MDA1986, DR081-1); breast cancer cell line (Hs578T), and non-malignant human cell line (MRC5)]. This led to the discovery of 15 withanolides, with IC50values in the range of 0.067−17.4 μM, including withaferin A, withaferin A 4,27-diacetate, 27-O-glucopyranosylwithaferin A, withalongolide H, withalongolide C, withalongolide A, withalongolide A 4,27-diacetate, withalongolide A 4,19,27-triacetate, withalongolide B, withalongolide B 4-acetate, withalongolide B 4,19-diacetate, withalongolide D, withalongolide E, withalongolide G, and 2,3-dihydrowithaferin A 3-O-sulfate. In order to update the growing literature on withanolides and their activities, we summarized the distribution, structural types, and antiproliferative activities for all published withanolides to date. The structure–activity relationship analysis (SARA) confirmed the importance of the presence of a ∆2-1-oxo-functionality in ring A, a 5β,6β-epoxy or 5α-chloro-6β-hydroxy grouping in ring B, and nine-carbon side chain with a lactone moiety for cytotoxic activity. Conversely, the SARA indicated that the –OH or –OR groups at C-4, 7, 11, 12, 14, 15, 16, 17, 18, 19, 20, 23, 24, 27, and 28 were not contributors to the observed antiproliferative activity within the systems analyzed.

Effects of Onosma dichroanthum Boiss. root extract on AGS human gastric cancer cell-line

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Mohammad Mostakhdem Hashemi ◽  
Majid Marjani ◽  
Nahid Poursharifi ◽  
Abdoljalal Marjani
Keyword(s):  
Gastric Cancer ◽  
Cell Line ◽  
Cancer Cell ◽  
Incubation Time ◽  
Gastric Cancer Cell ◽  
Gastric Cancer Cell Line ◽  
Cancer Cell Line ◽  
Root Extract ◽  
Human Gastric Cancer

Abstract One of the cancer-related deaths is gastric cancer in this area. Onosma dichroanthum Boiss. roots have been used as an antiseptic and anti-inflammatory for wound healing treatment. The aim of this study was to assess the in vitro cytotoxic and anticancer effects of O. dichroanthum Boiss. roots from the Golestan province of Iran. After identification of the taxonomical effect of O. dichroanthum Boiss., different concentrations of the hydroalcoholic root extract were used. Three different time periods (24, 48, and 72 h) were used to treat AGS gastric cancer and L-929 normal fibroblasts cell lines. The evaluation of different concentrations of root extract was performed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The 48 h treatment affected cell survival, while the concentration of 64 μg/mL was determined as IC50 concentrations at 48 h incubation time. The 48 h incubation time with 64 μg/mL showed the best effectiveness on cancerous cell-line while being safe for normal cell-line. Our results show that O. dichroanthum Boiss. roots extract may have cytotoxic and safe effects on gastric cancer cell-line and normal cells in 48 h treatment periods, respectively. The results indicated the O. dichroanthum Boiss. may be as an effective anticancer agent (gastric cancer).

scholarly journals In vitro Cytotoxic Activity of Sesamin Isolated from Vismia baccifera var. dealbata Triana & Planch (Guttiferae) Collected from Venezuela

2008 ◽  
Vol 3 (10) ◽  
pp. 1934578X0800301 ◽  
Author(s):  
Fabiola Salas ◽  
Janne Rojas ◽  
Antonio Morales ◽  
Maria E. Ramos-Nino ◽  
Nelida G. Colmenares
Keyword(s):  
Lung Cancer ◽  
Cytotoxic Activity ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Line ◽  
Lung Cancer Cell Line ◽  
Cytostatic Activity ◽  
Lung Cancer Cell

Sesamin extracted from Vismia baccifera var. dealbata was demonstrated to have cytostatic activity on the cancer cell lines tested, particularly the lung cancer cell line, with an IC50 of 1 g/L.

scholarly journals PHYTOCHEMICAL ANALYSIS, LIQUID CHROMATOGRAPHY, AND MASS SPECTROSCOPY AND IN VITRO ANTICANCER ACTIVITY OF ANNONA SQUAMOSA SEEDS LINN.

2018 ◽  
Vol 11 (3) ◽  
pp. 101
Author(s):  
Shuchi Dave Mehta ◽  
Sarvesh Paliwal
Keyword(s):  
Liquid Chromatography ◽  
Anticancer Activity ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Mass Spectroscopy ◽  
Cancer Cell Line ◽  
Phytochemical Analysis ◽  
Annona Squamosa

 Objective: The objective of the present study is to evaluate in vitro anticancer property and phytochemical analysis using liquid chromatography and mass spectroscopy (LCMS) method of hydroalcoholic extract of seeds of Annona squamosa (AS) Linn. Seeds of AS Linn. are traditional medicine treating various diseases and have shown anticancer activity. Due to lack of survival benefit, cancer is a deadly global disease.Method: The anticancer activity was evaluated using the sulforhodamine B assay method on five cancer cell lines: Breast cancer cell line, cervix cancer cell line (SiHa), colon cancer cell line (HT)-29, liver cancer cell line, and ovary cancer cell line (Ovcar). The phytochemical analysis was performed using LCMS method.Result: The phytochemical characterization was done using LCMS method which showed 15 different molecular weight compounds. The extract showed an average in vitro anticancer activity at a concentration of 100 μg/ml against all cancer cell lines. The best activity was observed against Ovcar-5 cell line (69.72) and was also significant against HT and SiHa cell lines.Conclusion: The phytochemical analysis showed the wide range of phenols and flavonoid which are showing potent anticancer activity of AS seeds.

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